. . () .

, S, .

. . . .

29. .

22

. . (), , , . , . .

- (S). .

k1 k3( kcat) + S ES E + P

k2

. . .

23

. . .

3036.

. . . . , , .

24

. , - . - . . .

:

1. ( reduce = )

.

.

.

NADH H+

.

25

+ NAD+ + NADH H+

.

22.

22 22 2

2.

.

.

:1.1. CHOH,

1.2. -

1.3. CH-CH-

1.4. CH-NH2 1.5. -CH-NH-

1.6. NADH, NADPH

1.10.

1.11 H2O21.12.

1.14.

26

1.7.

1.8.

1.9.

1.15

1.16.

1.17 CH2-

. 1.1

, -CHOH,

: 1.1.1. NAD+ NADP+,

1.1.2.

11.3. O2

1.1.99.

2.

.

.

.

-CH3, -COOH,

CO-R, NH2 .

, , -

27

, .

(),

GTP.

:

.

,

:2.1. (-, -,

-)

2.2.

2.3.

2.4.

2.5.

2.6.

2.7. ()

2.8.

28

+ +

3.

. ,

.

CO-

.

R-CO-O-R.

.

(C-O-PO3H2).

(-O-PO2H--).

.

29

DN, RN

DNA, RNA .

3.1

(, , ,

)

,

:3.1 (, ,

, )

3.2 -

3.3

3.4.

3.5. C-N ( )

3.6.

3.7 . C-C

3.8. C-X, P-X

3.9. P-N

3.10. C-P

4.

.

CO2

(-COOH)

30

. (-CH2-CH2-COOH

-CH=CH2 + CO2 ).

.

,

:4.1. C-C 4.2. C-O 4.3. C-N

4.4. C-S 4.5. C-X 4.6. P-O

4.99

5.

.

, cis- trans- ,

.

cis- trans-

:5.1. ,

5.2. cis-trans

31

C = CH H

R1 R2

C = CH

HR1

R2C = C

H H

R1 R2

C = CC = CH H

R1 R2

C = CH

HR1

R2C = CC = C

H

HR1

R2

CR3

R1H R2 C

R3

R1R2 HC

R3

R1H R2C

R3

R1H R2 C

R3

R1R2 HC

R3

R1R2 H

5.3. (

C=C, S-S,)

5.4. ( ,

, ..)

5.5.

6.

.

, ATP,

, GTP, .

,

:

6.1. C-O 6.2. C-S

6.3. C-N 6.4. C-C

6.5. -

:

32

1.

. :

NAD+ : --NADP+ : ---

NAD+ :

S2 + NAD+ S + DH + H+

S2 .

NADP+

S2 + NAD+ S + DH + H+

. : FMN - FAD

S2 + FMN+ S + FMNH2 S2 + FAD+ S + FADH2

. . . . . ( C)

2.

33

. P ( ): : , , (), ()

. (CoA) : (COCH3). : . : (CH3). ( , c):

: (CH3), (-CH2OH), (-CHO)

. ( ) : (COOH). CO2

. ( 1 ): . ( B6) :

(2) (COOH).

3. , ,

12 .

.

,

34

.

.. -

.

Enzyme

Commission EC

. , 1.1.1.1

.

1 : .

.

1.1. :

1.1.1.

NAD NADP

.

1.1.1.1

.

35

1. pH

. , - , . , pH . pH . pH . , pH . pH pH .

31. pH

36

pH

pH

, . : pH , pH , .

, , , , , , . (pH 4-6) : . . . . . . ( pH 5-7) . , .

37

(pH 10) . , , , , . , . , . . .

2.

, . . , . 50C. . 90C 60 70C .

38

3265.

6- (G6PD) . . . . . , G6PD.

3. .

39

=C

t

=C

t =

C

t

. , , .

33.

. .

4. .

32.

40

(mM)0 50 100 150 200 250 300 350

120

100

80

60

40

20

0

%

(mM)0 50 100 150 200 250 300 350

120

100

80

60

40

20

0

%

. , . , . S.

3436. Michaelis-Menten

Michaelis-Menten. . Michaelis :

[S] V = Vmax Km = k2 + kcat / k1 [S] + Km

, Vmax, ,

41

- . Vmax . V=Vmax/2 m, m=[S]. , Km( Michaelis), V=Vmax/2. m . . m . Vmax m . Km Vmax ( pH, , , ). Vmax/Km. . Vmax , kcat/Km . , . []=[S] (Vmax) S.

42

kcat[ES] E + P

([E]=[ES])

Vmax = kcat [ES] Vmax = kcat [E] kcat = Vmax/ [E]

kcat ( min sec) . mole/L.sec kcat sec-1, sec sec . O kcat/Km Vmax/Km kcat/Km = (Vmax/Km)/[E].

. .

Lineweaver-Burk , 1/v 1/S. , Km Vmax.

3536. Lineweaver-Burk

43

Km

, . , Km Km.

. . Km ( ) . .

5. .

44

CH3CH2OH + NAD CH3CHO + NADH2CH3CH2OH + NAD CH3CHO + NADH2

. . . .

. .

. . , - , , . .

, ()

+ , : H , , .

45

. ,

. . , , . , Km Vmax.

36. . : , : , :

.

. ,

. Vmax Km .

46

YY

A A

E

E

E

YY

A A

E

E

E

YY

A A

E

E

E

37.

.

. Vmax () Km ( ).

. ,

- S + I ESI . Km Vmax.

38.

, . .

47

A

A

A

A

A

A

A

AY

Y

Y

Y

Y

Y

YY

Y

YY

YY

Y

YY

.

. . (feedback inhibition), .

, L- L-, , L- 2 .

1 2 3 4 5L- L-

.

. . .

Mg2+, Mn2+, Ca2+, Ni2+, Zn2+ . .

48

. . , , . .

. . , . . . . . . .

49

.

39 65, 66.

.

1- , , . (). . glu342 Lys ( ), Lys342 Glu . .

50

.

, (Mevinolin,

Mevastatin, Lovastatin) 3--3-

--CoA (HMG-CoA )

. ,

.

. , . .

6.

,

51

. , , Vmax = kcat . [E]

40.

41. .

52

0 t

[P]

0 t

[P]

[S]

v

[S]

v

. , . () . ( pH).

. . , . . .

.

53

Vmax.

, , . . .

. . . . . , .

54

S PE

S PE

= = [P] [S]t t = = [P] [S]

t t

, . NAD NADP. NADH2 NADPH2 340 nm . FAD. 450 nm FADH2.

42. NAD FAD.

55

P1 P2E2

S P1E1

P1 P2E2

P1 P2E2

S P1E1

(nm)

(nm)

o (mole) . , (=mol / ) . ( pH ), . . (International Unit, IU, Unit, U).

56

COOH

H C NH2CH3

COOH

C O

CH3

COOH

H C NH2CH2CH2COOH

COOH

C O

CH2CH2COOH

+ +GPTCOOH

H C NH2CH3

COOH

H C NH2CH3

COOH

C O

CH3

COOH

C O

CH3

COOH

H C NH2CH2CH2COOH

COOH

H C NH2CH2CH2COOH

COOH

H C NH2CH2CH2COOH

COOH

C O

CH2CH2COOH

COOH

C O

CH2CH2COOH

+ +GPT

+ - + + - +

COOH

C O

CH3

COOH

H C OH

CH3

+ NADH H + + NAD+

COOH

C O

CH3

COOH

C O

CH3

COOH

H C OH

CH3

COOH

H C OH

CH3

+ NADH H + + NAD+

+ NAD + + NAD+340

+ NAD + + NAD+340

340 nm

Katal. katal mole .

mg katal kg . . .

. mole katal mole .

. Km Vmax . . .

57

, .

.

. , , , . . .

. , SDS . 30 . , .

, .

58

.

, , , , .

. . (heart), 4. (muscle), 4. 3, 22, 3. 4 4 .

. . 1:

. 2-5: . 2: , 3: , 4: , 5: .

(lactate dehydrogenase, LDH) . LDH . .

59

43. (CPK), (LDH) - (DH) .

. , . . .

60

, . . , .

. ,

. pH 5,

44 .

, ,

(, ,

) .

.

.

, -

61

-

.

-

-

.

44.

, pH, , , , .

.

, . ( ). . .

62

, . . .. , , . pH, . .

63